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Pinocchio di Ancona, Italy

Virology Laboratory

Pinocchio di Ancona, Italy
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News Article | December 2, 2016
Site: www.chromatographytechniques.com

Researchers from the University of São Paulo (USP) at Ribeirão Preto in Brazil have developed a platform that analyzes clinical samples from patients to diagnose infection by 416 viruses found in the world's tropical regions. According to its creators, the tool can be used by reference laboratories such as Adolfo Lutz Institute, Oswaldo Cruz Foundation (Fiocruz) and Evandro Chagas Institute in Brazil to assist epidemiological surveillance by detecting pathogens with the potential to cause epidemics in humans. Results of the research project, which was coordinated by Victor Hugo Aquino, a professor at the University of São Paulo's Ribeirão Preto Pharmaceutical School (FCFRP-USP), and supported by FAPESP, have been published in PLoS Neglected Tropical Diseases. "The number of patients with suspected dengue, Zika or chikungunya infection will increase when summer arrives," said Aquino, lead author of the article. "Conventional methods are frequently unable to confirm diagnosis of these diseases, so we don't know which viruses are circulating." In his view, if a tool like this had been available when Zika began circulating in Brazil, it might have been possible to restrict its spread to the initial outbreak location. "We took a long time to realize an epidemic was under way because no one was thinking of Zika at the time," he said. In addition to the pathogens, the platform detects others that as yet have been identified only sporadically but could become epidemics. Examples include Mayaro, an alphavirus related to chikungunya that is transmitted by wild mosquitoes such as Haemagogus janthinomys, and Oropouche, which to date has caused epidemics confined to riverine communities in the Amazon region and is transmitted mainly by midges of the species Culicoides paraensis. "There are several other viruses that haven't yet caused problems in humans but may do so one day," Aquino said. "They're evolving all the time, and with the degradation of natural environments infectious agents once confined to natural niches could spread farther afield." Although the platform is designed above all to detect pathogens transmitted by arthropods such as mosquitoes and ticks, it can also diagnose infectious agents transmitted by small mammals, like hantavirus. Aquino explained that the selection encompasses all viruses occurring in tropical regions with DNA sequences deposited in GenBank, a public database maintained by the National Center for Biotechnology Information (NCBI), which is part of the United States National Library of Medicine (NLM). The platform consists of a DNA microarray slide with eight identical sub-arrays containing viral probes replicated at least three times to complete the array with 15,000 probes. Each probe contains the sequences for 60 nucleotides that are complementary to the genomes of the viruses to be detected. According to Aquino, the sequences were mounted on the basis of information from GenBank using bioinformatics. "If a blood sample contains one of the 416 viruses included on the microchip, the pathogen's genome will bind with one of the probes to produce a marker that can be detected by a scanner," Aquino said. The device that reads the results is the same as that used in microarray assays for the analysis of gene expression. "Initially, the test will not be for the entire population because of high cost," Aquino said. "It will be used on patients with suspected dengue, Zika or other febrile diseases whose diagnosis isn't confirmed by conventional methods." The methodology was validated using 20 viruses available at FCFRP-USP's Virology Laboratory. The validation tests did not point to cross-hybridization, which produces a positive result for more than one infectious agent and hinders correct identification of single viruses. Nevertheless, the method proved effective to diagnose cases of co-infection, such as when the same patient has been infected by both Zika and dengue.


News Article | December 2, 2016
Site: www.rdmag.com

Researchers from the University of São Paulo (USP) at Ribeirão Preto in Brazil have developed a platform that analyzes clinical samples from patients to diagnose infection by 416 viruses found in the world's tropical regions. According to its creators, the tool can be used by reference laboratories such as Adolfo Lutz Institute, Oswaldo Cruz Foundation (Fiocruz) and Evandro Chagas Institute in Brazil to assist epidemiological surveillance by detecting pathogens with the potential to cause epidemics in humans. Results of the research project, which was coordinated by Victor Hugo Aquino, a professor at the University of São Paulo's Ribeirão Preto Pharmaceutical School (FCFRP-USP), and supported by FAPESP, have been published in PLoS Neglected Tropical Diseases. "The number of patients with suspected dengue, Zika or chikungunya infection will increase when summer arrives," said Aquino, lead author of the article. "Conventional methods are frequently unable to confirm diagnosis of these diseases, so we don't know which viruses are circulating." In his view, if a tool like this had been available when Zika began circulating in Brazil, it might have been possible to restrict its spread to the initial outbreak location. "We took a long time to realize an epidemic was under way because no one was thinking of Zika at the time," he said. In addition to the pathogens, the platform detects others that as yet have been identified only sporadically but could become epidemics. Examples include Mayaro, an alphavirus related to chikungunya that is transmitted by wild mosquitoes such as Haemagogus janthinomys, and Oropouche, which to date has caused epidemics confined to riverine communities in the Amazon region and is transmitted mainly by midges of the species Culicoides paraensis. "There are several other viruses that haven't yet caused problems in humans but may do so one day," Aquino said. "They're evolving all the time, and with the degradation of natural environments infectious agents once confined to natural niches could spread farther afield." Although the platform is designed above all to detect pathogens transmitted by arthropods such as mosquitoes and ticks, it can also diagnose infectious agents transmitted by small mammals, like hantavirus. Aquino explained that the selection encompasses all viruses occurring in tropical regions with DNA sequences deposited in GenBank, a public database maintained by the National Center for Biotechnology Information (NCBI), which is part of the United States National Library of Medicine (NLM). The platform consists of a DNA microarray slide with eight identical sub-arrays containing viral probes replicated at least three times to complete the array with 15,000 probes. Each probe contains the sequences for 60 nucleotides that are complementary to the genomes of the viruses to be detected. According to Aquino, the sequences were mounted on the basis of information from GenBank using bioinformatics. "If a blood sample contains one of the 416 viruses included on the microchip, the pathogen's genome will bind with one of the probes to produce a marker that can be detected by a scanner," Aquino said. The device that reads the results is the same as that used in microarray assays for the analysis of gene expression. "Initially, the test will not be for the entire population because of high cost," Aquino said. "It will be used on patients with suspected dengue, Zika or other febrile diseases whose diagnosis isn't confirmed by conventional methods." The methodology was validated using 20 viruses available at FCFRP-USP's Virology Laboratory. The validation tests did not point to cross-hybridization, which produces a positive result for more than one infectious agent and hinders correct identification of single viruses. Nevertheless, the method proved effective to diagnose cases of co-infection, such as when the same patient has been infected by both Zika and dengue.


News Article | December 1, 2016
Site: www.eurekalert.org

Researchers from the University of São Paulo have produced a tool for reference labs to assist epidemiological surveillance by analyzing clinical samples from patients with suspected infection by dengue, Zika, chikungunya and many other viral agent Researchers from the University of São Paulo (USP) at Ribeirão Preto in Brazil have developed a platform that analyzes clinical samples from patients to diagnose infection by 416 viruses found in the world's tropical regions. According to its creators, the tool can be used by reference laboratories such as Adolfo Lutz Institute, Oswaldo Cruz Foundation (Fiocruz) and Evandro Chagas Institute in Brazil to assist epidemiological surveillance by detecting pathogens with the potential to cause epidemics in humans. Results of the research project, which was coordinated by Victor Hugo Aquino, a professor at the University of São Paulo's Ribeirão Preto Pharmaceutical School (FCFRP-USP), and supported by FAPESP, have been published in PLoS Neglected Tropical Diseases. "The number of patients with suspected dengue, Zika or chikungunya infection will increase when summer arrives," said Aquino, lead author of the article. "Conventional methods are frequently unable to confirm diagnosis of these diseases, so we don't know which viruses are circulating." In his view, if a tool like this had been available when Zika began circulating in Brazil, it might have been possible to restrict its spread to the initial outbreak location. "We took a long time to realize an epidemic was under way because no one was thinking of Zika at the time," he said. In addition to the pathogens, the platform detects others that as yet have been identified only sporadically but could become epidemics. Examples include Mayaro, an alphavirus related to chikungunya that is transmitted by wild mosquitoes such as Haemagogus janthinomys, and Oropouche, which to date has caused epidemics confined to riverine communities in the Amazon region and is transmitted mainly by midges of the species Culicoides paraensis. "There are several other viruses that haven't yet caused problems in humans but may do so one day," Aquino said. "They're evolving all the time, and with the degradation of natural environments infectious agents once confined to natural niches could spread farther afield." Although the platform is designed above all to detect pathogens transmitted by arthropods such as mosquitoes and ticks, it can also diagnose infectious agents transmitted by small mammals, like hantavirus. Aquino explained that the selection encompasses all viruses occurring in tropical regions with DNA sequences deposited in GenBank, a public database maintained by the National Center for Biotechnology Information (NCBI), which is part of the United States National Library of Medicine (NLM). The platform consists of a DNA microarray slide with eight identical sub-arrays containing viral probes replicated at least three times to complete the array with 15,000 probes. Each probe contains the sequences for 60 nucleotides that are complementary to the genomes of the viruses to be detected. According to Aquino, the sequences were mounted on the basis of information from GenBank using bioinformatics. "If a blood sample contains one of the 416 viruses included on the microchip, the pathogen's genome will bind with one of the probes to produce a marker that can be detected by a scanner," Aquino said. The device that reads the results is the same as that used in microarray assays for the analysis of gene expression. "Initially, the test will not be for the entire population because of high cost," Aquino said. "It will be used on patients with suspected dengue, Zika or other febrile diseases whose diagnosis isn't confirmed by conventional methods." The methodology was validated using 20 viruses available at FCFRP-USP's Virology Laboratory. The validation tests did not point to cross-hybridization, which produces a positive result for more than one infectious agent and hinders correct identification of single viruses. Nevertheless, the method proved effective to diagnose cases of co-infection, such as when the same patient has been infected by both Zika and dengue.


Aghokeng A.F.,Virology Laboratory | Aghokeng A.F.,Montpellier University | Kouanfack C.,Central Hospital | Laurent C.,Montpellier University | And 8 more authors.
AIDS | Year: 2011

OBJECTIVES:: To evaluate the frequency and progression over time of the WHO-defined transmitted HIV-1 drug resistance mutations (DRMs) among antiretroviral treatment (ART)-naive HIV-1-infected patients in Cameroon. DESIGN:: We analyzed HIV-1 DRM data generated from 369 ART-naive individuals consecutively recruited between 1996 and 2007 in urban and rural areas in Cameroon. METHODS:: HIV-1 drug resistance genotyping was performed in the pol gene using plasma samples and surveillance DRMs were identified using the 2009 WHO-DRM list. RESULTS:: We observed in Yaounde, the capital city, an increasing prevalence of DRMs over time: 0.0% (none of 61 participants) in 1996-1999; 1.9% (one of 53 participants) in 2001; 4.1% (two of 49 participants) in 2002; and 12.3% (10 of 81 participants) in 2007. In the rural areas with more recently implemented ART programs, we found DRMs in six of 125 (4.8%) ART-naive individuals recruited in 2006-2007. DRMs identified in both areas included resistance mutations to protease inhibitors, nucleoside reverse transcriptase inhibitors (NRTIs) and non-NRTIs (NNRTIs) that might impair the efficacy of available first-line and second-line treatments. CONCLUSION:: This report showed an increase in transmitted DRMs in areas where antiretroviral drugs were introduced earlier, although other factors such as natural viral polymorphisms and acquired DRMs through exposure to antiretroviral cannot be totally excluded. Further surveillances are needed to confirm this evolution and inform public health policies on adequate actions to help limit the selection and transmission of drug-resistant HIV, while scaling up access to ART in developing countries. © 2011 Wolters Kluwer Health Lippincott Williams & Wilkins.


Hubschen J.M.,Institute of Immunology | Mbah P.O.,District Hospital Bali | Forbi J.C.,Institute of Immunology | Forbi J.C.,Virology Laboratory | And 4 more authors.
Clinical Microbiology and Infection | Year: 2011

To further investigate the genetic diversity of hepatitis B virus (HBV) genotype A in Africa, we analysed 263 HBV strains from Nigeria (n = 163) and Cameroon (n = 100). Phylogenetic analysis of S fragment sequences attributed 175 strains (66.5%) to genotype E and 88 (33.5%) to genotype A. In Cameroon, genotype A strains were the most prevalent (79/100, 79.0%), whereas, in Nigeria, genotype E was highly dominant (154/163, 94.5%). The genotype A strains grouped with reference strains of subgenotype A3 (n = 8), the provisional subgenotype A5 (n = 43), a recently reported new variant from Rwanda (n = 35), or as outliers (n = 2). Ten complete genome sequences obtained from strains that clustered with the new variant from Rwanda formed a separate group supported by a bootstrap value of 96. The between- group distance to other potential or recognized subgenotypes of genotype A was at least 3.81%. Thus, the new group of strains could be considered as a new subgenotype of HBV genotype A, tentatively named 'A7'. Interestingly, the 'A7' strains from Rwanda and Cameroon showed an interspersed clustering, but essentially no other (sub)genotypes were shared between the two countries, suggesting that 'A7' may have evolved in a yet unknown place and may have only relatively recently spread to Rwanda and Western Cameroon. Strains attributed to provisional subgenotype A5 were found for the first time in Cameroon (n = 36) and Central Nigeria (n = 2), indicating that A5 is more widespread than previously thought. © 2010 The Authors. Journal Compilation © 2010 European Society of Clinical Microbiology and Infectious Diseases.


Hoarau J.-J.,University of Reunion Island | Krejbich-Trotot P.,University of Reunion Island | Jaffar-Bandjee M.-C.,University of Reunion Island | Jaffar-Bandjee M.-C.,Virology Laboratory | And 5 more authors.
CNS and Neurological Disorders - Drug Targets | Year: 2011

Innate immunity is an arsenal of molecules and receptors expressed by professional phagocytes, glial cells and neurons and involved in host defence and clearance of toxic and dangerous cell debris. However, any uncontrolled innate immune responses within the central nervous system (CNS) are widely recognized as playing a major role in the development of autoimmune disorders and neurodegeneration, with multiple sclerosis (MS) and Alzheimer's diseases (AD) being primary examples. Critically, neuroimmune regulatory proteins (NIReg) may control the adverse immune responses in health and diseases. NIRegs are found mainly on neurons, glia, endothelia and ependymal cells and include GPI-anchored molecules (CD24, CD90, complement regulators CD55 and CD59), molecules of the immunoglobulin superfamily (siglec CD22, Siglec 10, CD200, ICAM-5) and others (CD47, fractalkine, TAM receptor tyrosine kinase and complement C3a and factor H). These regulators modulate the innate immune response in the CNS and for instance critically control the level of phagocytosis and inflammation engaged by resident microglia and infiltrating immune cells. Others will sequester and neutralize proinflammatory molecules such as HMGB1 and DNA. Moreover, some NIRegs can instigate the recruitment of stem cells to mediate tissue repair. In the absence of these regulators, when neurons die by apoptosis, become infected or damaged, microglia and infiltrating immune cells are free to cause injury and an adverse inflammatory response in acute and chronic settings. The therapeutic applications of NIRegs should be exploited given their natural and selective healing properties. © 2011 Bentham Science Publishers.


Rottenstreich A.,Rambam Health Care Campus | Rottenstreich A.,Technion - Israel Institute of Technology | Oz Z.K.,Virology Laboratory | Oren I.,Rambam Health Care Campus | Oren I.,Technion - Israel Institute of Technology
Diagnostic Microbiology and Infectious Disease | Year: 2014

Varicella zoster virus (VZV) like other alphaherpes viruses stays latent after its primary infection. During its reactivation, it can infect the central nervous system (CNS) causing a variety of clinical presentations. Using polymerase chain reaction (PCR) for detection of VZV DNA in cerebrospinal fluid (CSF), it is now recognized in some series as the most common causative agent of viral CNS infection. We aimed to investigate in our study the correlation between VZV viral load in the CSF and the clinical course of its infection, using quantitative real-time PCR. For this purpose, we examined 56 specimens of consecutive patients with positive CSF for VZV DNA in a qualitative test, with a clinical picture of meningitis or encephalitis collected over 10. years in Rambam medical center. We found a significant correlation between VZV viral load and the severity and duration of neurological disease. We believe that using quantitative measurement of VZV DNA in the CSF, could serve as a prognostic marker which would influence treatment decisions. © 2014 Elsevier Inc.


Pyndiah M.N.,Virology Laboratory
Médecine tropicale : revue du Corps de santé colonial | Year: 2012

During the chikungunya outbreak of 2005 - 2006, the only laboratory facilities available in Mauritius were virus isolation in cell culture tubes and serology. The laboratory was submerged with large numbers of blood samples. Comparative isolation was made in human embryonic lung (HEL) and VERO cells grown in 96-well plate. Culture on HEL cells was found to be more sensitive and presence of cytopathic effect (CPE) was observed earlier than in VERO cells. Out of the 18 300 blood samples inoculated on HEL, 11 165 were positive. This virus isolation method was of great help for the surveillance and control of the vectors. In cases of an outbreak a cheap, rapid and simple method of isolating chikungunya virus is described.


Das T.,University of Reunion Island | Hoarau J.J.,University of Reunion Island | Bandjee M.C.J.,University of Reunion Island | Bandjee M.C.J.,Virology laboratory | And 2 more authors.
Journal of General Virology | Year: 2015

Chikungunya virus (CHIKV) has recently affected millions of people in the Indian Ocean, with rare cases of encephalopathy and encephalitis occurring in neonates. In the study described herein, the capacity of mouse brain cells to control infection through innate immune antiviral responses was assessed. In vitro, CHIKV principally infected a subpopulation of mouse GFAP+ primary astrocytes. Oligodendrocytes and neurons could also be infected. An innate immune response was engaged by CHIKV-infected astrocytes with elevated expression of mRNAs for IFN-α−β, inflammatory cytokines (e.g. IL-1β, IL-12, IL-10, IL-24) and proapoptotic factors (e.g. TNF-α, FasL, Lymphotoxin B). Programmed cell death through the intrinsic caspase-9 pathway was observed by immunofluorescence in infected astrocytes and neurons but not in oligodendrocytes. Interestingly, microglia did not replicate CHIKV but responded by elevated mitogen-activated protein kinase (MAPK) activity. Intracerebroventricular injection of CHIKV in neonate mice led to the infection of astrocytes. The astrogliosis response was accompanied by a dendritic CD206+ cell mobilization restricted to the site of infection. The results of this study support the paradigm that a multifaceted innate immune response can be mobilized by both professional immune and glial cells to control CHIKV neuroinfection events in neonates. © 2015 The Authors.


Houhou N.,Virology Laboratory | Choquet C.,Hopital Bichat Claude Bernard
Clinical Microbiology and Infection | Year: 2014

We reviewed 80 adult cases of measles seen in a Parisian hospital during the French 2010-2011 outbreak. Fifty per cent had at least one complication: pneumonia and hepatitis were the most frequent. Forty per cent of hospitalized cases did not have any complications, suggesting clinically poor tolerance of measles in adults. The outcome was always favourable. Subjects were younger, were more often French nationals and had a higher socio-economic status than the overall population. This report suggests that immunity resulting from natural disease in patients from an area where the disease is endemic is protective in the long term. © 2013 European Society of Clinical Microbiology and Infectious Diseases.

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